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Phospho-DRP1 (S616) Recombinant Rabbit Monoclonal Antibody [PSH06-77]

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Specification

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Catalog# HA722760

Phospho-DRP1 (S616) Recombinant Rabbit Monoclonal Antibody [PSH06-77]

Application

  • WB

  • IHC-P

  • IF-Tissue

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-DRP1 (S616) Recombinant Rabbit Monoclonal Antibody [PSH06-77]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser616 of human DRP1.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Tissue, IF-Cell, FC

Molecular Weight

Predicted band size: 82 kDa

Positive Control

HeLa cell lysate, HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 100ng/mL nocodazole for 18 hours cell lysate, HeLa, C2C12 cell lysate, PC-12 cell lysate, mouse brain tissue, Jurkat cell lysate, HEK-293 cell lysate, Human brain tissue lysate, Mouse brain tissue lysate, Mouse heart tissue lysate, Rat brain tissue lysate, Rat heart tissue lysate, rat brain tissue, PC-12.

Conjugation

unconjugated

Clone Number

PSH06-77

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IHC-P

  • 1:10,000

  • IF-Tissue

  • 1:500-1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Dynamin-1-like protein is a GTPase that regulates mitochondrial fission. In humans, dynamin-1-like protein, which is typically referred to as dynamin-related protein 1 (Drp1), is encoded by the DNM1L gene and is part of the dynamin superfamily (DSP) family of proteins. Mitochondria routinely undergo fission and fusion events that maintain a dynamic reticular network. Drp1 is a fundamental component of mitochondrial fission. Indeed, Drp1 deficient neurons have large, strongly interconnected mitochondria due to dysfunctional fission machinery. Fission helps facilitate mitophagy, which is the breakdown and recycling of damaged mitochondria. Dysfunction in the DRP activity may result in mutated DNA or malfunctioning proteins diffusing throughout the mitochondrial system. In addition, fission results in fragmented mitochondria more capable of producing of reactive oxygen species, which can disrupt normal biochemical processes inside of cells. ROS can be formed from incomplete transfer of electrons through the electron transport chain. Furthermore, fission influences calcium flux within the cell, linking Drp1 to apoptosis and cancer.

Background References

1. Zeng X et al. Activated Drp1 regulates p62-mediated autophagic flux and aggravates inflammation in cerebral ischemia-reperfusion via the ROS-RIP1/RIP3-exosome axis. Mil Med Res. 2022 May

2. Jin JY et al. Drp1-dependent mitochondrial fission in cardiovascular disease. Acta Pharmacol Sin. 2021 May

Subcellular Location

Cytoplasm, cytosol, Golgi apparatus, Endomembrane system, Mitochondrion outer membrane, Peroxisome, Membrane, clathrin-coated pit, Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane.

UNIPROT

SwissProt: O00429 Human

SwissProt: Q8K1M6 Mouse

SwissProt: O35303 Rat

Synonyms

DLP1 antibody

dnm1l antibody

DNM1L_HUMAN antibody

Dnm1p/Vps1p-like protein antibody

dnml1 antibody

DRP1 antibody

DVLP antibody

Dymple antibody

Dynamin 1 like antibody

Dynamin family member proline-rich carboxyl-terminal domain less antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: NIH/3T3 treated with 100ng/mL nocodazole for 18 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: Lane 1-4 (left): 1 minute 30 seconds; ECL: K1801;<br />Exposure time: Lane 1-4 (right): 1 minute 40 seconds; ECL: K1802;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 100ng/mL Calyculin A for 30 minutes cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: NIH/3T3 treated with 100ng/mL nocodazole for 18 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 75 kDa

    Exposure time: Lane 1-4 (left): 1 minute 30 seconds; ECL: K1801;
    Exposure time: Lane 1-4 (right): 1 minute 40 seconds; ECL: K1802;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722760) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Flow cytometric analysis of HeLa cells labeling Phospho-DRP1 (S616).<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Phospho-DRP1 (S616).

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722760, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: C2C12 cell lysate<br />Lane 3: PC-12 cell lysate<br />Lane 4: HeLa cell lysate, the membrane blocked with phospho-peptide<br />Lane 5: C2C12 cell lysate, the membrane blocked with phospho-peptide<br />Lane 6: PC-12 cell lysate, the membrane blocked with phospho-peptide<br />Lane 7: HeLa cell lysate, the membrane blocked with non-phospho-peptide<br />Lane 8: C2C12 cell lysate, the membrane blocked with non-phospho-peptide<br />Lane 9: PC-12 cell lysate, the membrane blocked with non-phospho-peptide<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: C2C12 cell lysate
    Lane 3: PC-12 cell lysate
    Lane 4: HeLa cell lysate, the membrane blocked with phospho-peptide
    Lane 5: C2C12 cell lysate, the membrane blocked with phospho-peptide
    Lane 6: PC-12 cell lysate, the membrane blocked with phospho-peptide
    Lane 7: HeLa cell lysate, the membrane blocked with non-phospho-peptide
    Lane 8: C2C12 cell lysate, the membrane blocked with non-phospho-peptide
    Lane 9: PC-12 cell lysate, the membrane blocked with non-phospho-peptide

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 75 kDa

    Exposure time: 25 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722760) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/10,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    ☑ Cell treatment (CT)

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/10,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722760) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: Jurkat cell lysate<br />Lane 3: HEK-293 cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: PC-12 cell lysate<br />Lane 7: Human brain tissue lysate<br />Lane 8: Mouse brain tissue lysate<br />Lane 9: Mouse heart tissue lysate<br />Lane 10: Rat brain tissue lysate<br />Lane 11: Rat heart tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 82 kDa<br />Observed band size: 75 kDa<br /><br />Exposure time: 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-DRP1 (S616) on different lysates with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: HEK-293 cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: PC-12 cell lysate
    Lane 7: Human brain tissue lysate
    Lane 8: Mouse brain tissue lysate
    Lane 9: Mouse heart tissue lysate
    Lane 10: Rat brain tissue lysate
    Lane 11: Rat heart tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 82 kDa
    Observed band size: 75 kDa

    Exposure time: 20 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722760) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/10,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/10,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722760) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunocytochemistry analysis of HeLa cells labeling Phospho-DRP1 (S616) with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling Phospho-DRP1 (S616) with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of PC-12 cells labeling Phospho-DRP1 (S616) with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-DRP1 (S616) antibody (<a href="/products/HA722760" style="font-weight: bold;text-decoration: underline;">HA722760</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling Phospho-DRP1 (S616) with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-DRP1 (S616) antibody (HA722760) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

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